U.S. patent application number 12/668773 was filed with the patent office on 2010-07-29 for peptide whitening agents and cosmetic compositions comprising the same.
This patent application is currently assigned to CHANEL PARFUMS BEAUTE. Invention is credited to Stefan Honing, Gaelle Saintigny.
Application Number | 20100190721 12/668773 |
Document ID | / |
Family ID | 39832521 |
Filed Date | 2010-07-29 |
United States Patent
Application |
20100190721 |
Kind Code |
A1 |
Honing; Stefan ; et
al. |
July 29, 2010 |
PEPTIDE WHITENING AGENTS AND COSMETIC COMPOSITIONS COMPRISING THE
SAME
Abstract
The present invention relates to novel peptide compounds having
the following formula
[R.sub.1R.sub.2N--CH(Ra)--CO].sub.a-(AA.sub.1).sub.m-(AA.sub.2).sub.b--X.-
sub.1--X.sub.2--X.sub.3--Y.sub.1--Y.sub.2--Y.sub.3--X.sub.4-(AA.sub.3).sub-
.n--[NH--CH(Rb)--COOR.sub.3].sub.c. It also pertains to cosmetic
and dermatological compositions comprising one or more of these
peptide compounds and to their uses in the cosmetic whitening of
human skin and for the manufacture of a dermatological preparation
intended to depigment human skin.
Inventors: |
Honing; Stefan; (Euskirchem,
DE) ; Saintigny; Gaelle; (Paris, FR) |
Correspondence
Address: |
YOUNG & THOMPSON
209 Madison Street, Suite 500
Alexandria
VA
22314
US
|
Assignee: |
CHANEL PARFUMS BEAUTE
Neuilly Sur Seine France
FR
|
Family ID: |
39832521 |
Appl. No.: |
12/668773 |
Filed: |
June 16, 2008 |
PCT Filed: |
June 16, 2008 |
PCT NO: |
PCT/EP08/57558 |
371 Date: |
January 12, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60929818 |
Jul 13, 2007 |
|
|
|
Current U.S.
Class: |
514/18.8 ;
530/328 |
Current CPC
Class: |
A61Q 19/02 20130101;
A61K 8/64 20130101 |
Class at
Publication: |
514/15 ;
530/328 |
International
Class: |
A61K 38/08 20060101
A61K038/08; C07K 7/06 20060101 C07K007/06; A61Q 90/00 20090101
A61Q090/00 |
Claims
1-14. (canceled)
15. Compound having the following formula (I):
[R.sub.1R.sub.2N--CH(Ra)--CO].sub.a-(AA.sub.1).sub.m-(AA.sub.2).sub.b--X.-
sub.1--X.sub.2--X.sub.3--Y.sub.1--Y.sub.2--Y.sub.3--X.sub.4-(AA.sub.3).sub-
.n--[NH--CH(Rb)--COOR.sub.3].sub.c (I) wherein: R.sub.1, R.sub.2,
R.sub.3 each independently represent: H, a C.sub.2-C.sub.22
optionally hydroxylated and/or sulphured linear hydrocarbon group
which may be saturated or unsaturated, a C.sub.3-C.sub.22
optionally hydroxylated and/or sulphured branched or cyclic
hydrocarbon group which may be saturated or unsaturated, or a
biotin group wherein at least one of R.sub.1 and R.sub.2 may
alternatively be a protective group or a blocking group Ra and Rb
each independently represent: H; a C.sub.1-C.sub.4 linear alkyl
group; or a C.sub.3-C.sub.4 branched alkyl group, each alkyl group
optionally being substituted by one or more substituents selected
from the group consisting of: OH, SH, COOH, CONH.sub.2,
NH.sub.2--S--CH.sub.3, NH--C(NH)--NH.sub.2 or by a cyclic or
heterocyclic aromatic group, X.sub.1, X.sub.2 and X.sub.4 each
independently represent a polar amino acid group, X.sub.3 is a
small, preferably uncharged, amino acid group, Y.sub.1, Y.sub.2 and
Y.sub.3 each independently represents a hydrophobic amino acid
group with the proviso that at least one of Y.sub.1 and Y.sub.2 is
leucine or isoleucine, AA.sub.1, AA.sub.2 and AA.sub.3
independently designate amino acid groups, wherein the AA.sub.1 and
AA.sub.3 amino acid groups, respectively, may be the same or
different when m and n are more than 1, m and n independently
represent an integer ranging from 0 to 10, a, b and c are
independently 0 or 1, wherein all the amino acid groups may
independently be in a L, D or DL configuration, with the proviso
that when X.sub.1 is arginine (R) or lysine (K), then Y.sub.3 is
different from methionine (M).
16. Compound according to claim 15, wherein the polar amino acid
groups are selected from the group consisting of: asparagine (N),
serine (S), cysteine (C), glutamine (Q), aspartic acid (D),
glutamic acid (E), threonine (T), lysine (K), arginine (R),
histidine (H), tyrosine (Y) and tryptophan (W).
17. Compound according to claim 15, wherein the small amino acid
groups are selected from the group consisting of: proline (P),
asparagine (N), aspartic acid (D), threonine (T), valine (V),
cysteine (C), alanine (A), serine (S) and glycine (G).
18. Compound according to claim 15, wherein the hydrophobic amino
acid groups are selected from the group consisting of: alanine (A),
glycine (G), cysteine (C), threonine (T), valine (V), isoleucine
(I), leucine (L), lysine (K), methionine (M), histidine (H),
tyrosine (Y), tryptophan (W) and phenylalanine (F).
19. Compound according to claim 15, wherein the R.sub.1 and/or
R.sub.2 hydrocarbon groups are alkyl carbonyl groups having from 6
to 20 carbon atoms.
20. Compound according to claim 19, wherein the R.sub.1 and/or
R.sub.2 hydrocarbon groups are a palmitoyl group.
21. Compound according to claim 15, wherein AA.sub.2 designates a
polar amino acid group.
22. Compound according to claim 21, wherein the polar amino acid
group is negatively charged.
23. Compound according to claim 21, wherein AA.sub.2 is glutamic
acid (E).
24. Compound according to claim 15, wherein X.sub.4 is aspartic
acid (D) or glutamic acid (E)
25. Compound according to claim 15, wherein X.sub.1 is arginine
(R), aspartic acid (D) or glutamic acid (E)
26. Compound according to claim 15, wherein X.sub.3 is proline
(P).
27. Compound according to claim 15, wherein Y.sub.1 and/or Y.sub.2
and/or Y.sub.3 is valine (V), isoleucine (I) or leucine (L).
28. Compound according to claim 15, wherein Y.sub.1 is leucine (L),
methionine (M) or isoleucine.
29. Compound according to claim 15, wherein Y.sub.2 is leucine
(L).
30. Compound according to claim 15, wherein Y.sub.3 is valine (V),
leucine (L) or methionine (M).
31. Compound according to claim 15, wherein X.sub.2 is glutamine
(Q), glutamic acid (E) or asparagine (N).
32. Compound according to claim 15, wherein: b=1 and AA.sub.2 is
glutamic acid (E), X.sub.1 is arginine (R), aspartic acid (D) or
glutamic acid (E), X.sub.2 is glutamine (Q), glutamic acid (E) or
asparagine (N), X.sub.3 is proline (P), X.sub.4 is aspartic acid
(D) or glutamic acid (E), Y.sub.1 is leucine (L), methionine (M) or
isoleucine (I), Y.sub.2 is leucine (L), Y.sub.3 is valine (V),
leucine (L) or methionine (M), and either n=m=a=c=0, or n=m=c=0 and
a=1, with the proviso that when X.sub.1 is arginine (R), then
Y.sub.3 is different from methionine (M).
33. Compound according to claim 15, wherein: b=1 and AA.sub.2 is
glutamic acid (E), X.sub.1 is aspartic acid (D), X.sub.2 is
glutamic acid (E), X.sub.3 is proline (P), X.sub.4 is glutamic acid
(E), Y.sub.1 is leucine (L), Y.sub.2 is leucine (L), Y.sub.3 is
methionine (M), and either n=m=a=c=0, or n=m=c=0 and a=1.
34. Cosmetic or dermatological composition comprising at least one
peptide compound as described in claim 15, or a mixture thereof, in
a physiologically acceptable medium.
35. Cosmetic or dermatological composition comprising at least one
peptide compound as described in claim 33, or a mixture thereof, in
a physiologically acceptable medium.
36. Cosmetic method for whitening, bleaching or lightening human
skin, comprising topically applying onto human skin a cosmetic
composition according to claim 34.
Description
[0001] The present invention relates to novel peptide compounds
that are specifically recognized by heterotetrameric adaptor
protein complexes (AP) and more specifically by AP3 and AP2 adaptor
complexes having a key role in the transport of tyrosinase and thus
in melanogenesis. It also pertains to cosmetic compositions
comprising these compounds, to a cosmetic whitening method
comprising the topical application of such compositions onto human
skin and to the use of this composition for manufacturing a
dermatological preparation intended to depigment human skin.
[0002] Extensive research has been conducted over the past years to
extract or synthesize new whitening agents to be used in cosmetic
and dermatological compositions in order to alleviate skin
pigmentation defects such as senile lentigo or chloasma or to
satisfy consumers who are eager to have the lightest possible
complexion.
[0003] Various whitening agents acting on melanogenesis have been
proposed to this end. Among them, however, some only show a slight
melanogenesis-inhibiting effect failing to provide a sufficient
whitening effect, while others, which are more effective, have
proven not to be free of side effects on account of their toxicity
to human skin cells, rendering their use quite hazardous in
cosmetics. This toxicity arises from the fact that they interfere
with fundamental mechanisms of melanogenesis, by killing cells and
thus forcing the skin to eliminate them while producing toxins.
[0004] For instance, hydroquinone is a compound which is
particularly irritant and cytotoxic to melanocytes, and whose total
or partial replacement has been contemplated by many investigators.
In order to overcome the drawbacks mentioned above, it has been
suggested, for example, to use derivatives of active compounds such
as fatty acid esters or glycosyl ethers of hydroquinone as
depigmenting agents. Unfortunately, although more photostable and
less toxic than hydroquinone, these derivatives, such as arbutin,
are less active than hydroquinone to improve the aspect of
pigmentary defects.
[0005] Other substances have been sought which are not involved in
the mechanism of melanogenesis, but which act upstream on
tyrosinase by preventing its activation, and are consequently much
less toxic. Tyrosinase is regarded as the key enzyme in melanin
synthesis, and catalyses the first two reactions, the hydroxylation
of L-tyrosine to 3,4-dihydrophenylalanine (DOPA) and the oxidation
of DOPA to DOPAquinone. Kojic acid is commonly used as a
tyrosinase-activation inhibitor by complexing the copper present in
the active site of the enzyme. Unfortunately, this compound can
give rise to allergic reactions (Nakagawa et al, in Contact
Dermatitis, 1995). In addition, this compound is unstable in
solution, which somewhat complicates the manufacture of a
composition containing it.
[0006] Other whitening agents acting on tyrosinase activity are
plant extracts, the efficacy of which is not always satisfactory. A
possible explanation for the limited efficacy of these known
whitening agents is that they lack proper access to tyrosinase
inside the melanosomes. The melanosomal membrane is indeed a very
impervious structure which protects the cell from very toxic
metabolites (quinones) generated during melanogenesis.
[0007] Another approach to inhibit skin melanogenesis is to act on
the process of tyrosinase maturation. Tunicamycin has for instance
been proposed to this end, since this compound disturbs tyrosinase
glycosylation which is necessary for its maturation and for its
migration from the Golgi to the melanosomes. However, the
inhibition of glycosylation is unspecific and associated with a
number of side effects rendering the cosmetic use of this compound
unsuitable. Similarly, other agents have been proposed which are
able to modify the conformation of tyrosinase so as to lower its
stability and direct it to the degradation pathways (ER-associated
degradation or through the endosomal/lysosomal system). However,
the major drawback of this approach is that it may also interfere
with the maturation of other proteins and thus alter a number of
biological mechanisms other than melanogenesis. Again, this is not
conceivable for a cosmetic product.
[0008] Thus, there is still a need for an agent for bleaching human
skin which acts as effectively as, or even better than known
agents, but which does not exhibit their drawbacks, i.e. which is
non-irritant, non-toxic and/or non-allergenic to the skin and which
is stable in a composition.
[0009] To satisfy this need, this invention uses an alternative way
for inhibiting the pigmentation process which is not only safer but
also very effective since it acts upstream of melanosome formation,
well before the formation of its membrane. More specifically, the
novel compounds of this invention have been conceived so as to
interfere with the intracellular transport of tyrosinase to
melanosomes. The problem of accessibility of the active agents to
their biological target is thus overcome. Moreover, these compounds
have a rather specific activity against melanogenesis, since they
inhibit and/or reduce tyrosinase transport from the TGN (Trans
Golgi Network) to melanosomes. They are thus not likely to
interfere with any other biological process, which makes them well
suited for a cosmetic application.
[0010] Melanosomes differ from lysosomes by the presence of enzymes
involved in pigment synthesis, as for example tyrosinase.
Tyrosinase is only active in a maturing melanosome, but not in
other organelles of its biosynthetic pathway. Thus, the approach
chosen by the inventors is to use short peptides to block the
binding of tyrosinase to the cytosolic heterotetrameric adaptor
complexes (AP) and specifically to AP-2 and AP-3, which are a key
component of the intracellular sorting machinery (Honing et al.,
EMBO J., 17, n.degree. 5, 1304-1314, 1998 and Honing et al., Mol.
Cell., 2005, 18, 519-531). One of the key function of AP complexes
is to bind membrane proteins such as tyrosinase via short peptide
sequences located in their intracellular tail. By binding
cytoplasmic sorting signals on cargo proteins, AP complexes recruit
cargo to patches on donor membranes that bud to form vesicles or
tubules intended to fuse with target membranes (Bonifacio and
Traub, Annu. Rev. Biochem., 72, 395-447, 2003).
[0011] More specifically, it has been demonstrated that tyrosinase
has a high affinity interaction with AP-3. AP-3 is an adaptor
complex composed of the four subunits .delta.-adaptin,
.beta.3A-adaptin, the medium chain .mu.3A and the small chain
.sigma.3A. It was demonstrated (DELL'ANGELICA E C et al., EMBO J.,
16, 917-928, 1997) that the .mu.3 chain can interact with
tyrosine-based sorting signals and that AP-3 binds dileucine-based
signals, which are involved in the transport of tyrosinase to
melanosomes. Preventing the binding between AP3 and tyrosinase
should thus inhibit the incorporation of tyrosinase in the
formation of the melanosomes and thus melanogenesis.
[0012] The novel compounds of this invention are peptide compounds
which, to the best of the inventors's knowledge, have never been
described or used before in cosmetics.
[0013] The present invention thus pertains to a compound having the
following formula (I):
[R.sub.1R.sub.2N--CH(Ra)--CO].sub.a-(AA.sub.1).sub.m-(AA.sub.2).sub.b--X-
.sub.1--X.sub.2--X.sub.3--Y.sub.1--Y.sub.2--Y.sub.3--X.sub.4-(AA.sub.3).su-
b.n--[NH--CH(Rb)--COOR.sub.3].sub.c (I)
[0014] wherein:
[0015] R.sub.1, R.sub.2, R.sub.3 each independently represent:
[0016] H, [0017] a C.sub.2-C.sub.22 optionally hydroxylated and/or
sulphured linear hydrocarbon group which may be saturated or
unsaturated, [0018] a C.sub.3-C.sub.22 optionally hydroxylated
and/or sulphured branched or cyclic hydrocarbon group which may be
saturated or unsaturated, or [0019] a biotin group,
[0020] wherein at least one of R.sub.1 and R.sub.2 may
alternatively be a protective group or a blocking group
[0021] Ra and Rb each independently represent: H; a C.sub.1-C.sub.4
linear alkyl group; or a C.sub.3-C.sub.4 branched alkyl group, each
alkyl group optionally being substituted by one or more
substituents selected from the group consisting of: OH, SH, COOH,
CONH.sub.2, NH.sub.2--S--CH.sub.3, NH--C(NH)--NH.sub.2 or by a
cyclic or heterocyclic aromatic group,
[0022] X.sub.1, X.sub.2 and X.sub.4 each independently represent a
polar amino acid group,
[0023] X.sub.3 is a small, preferably uncharged, amino acid
group,
[0024] Y.sub.1, Y.sub.2 and Y.sub.3 each independently represents a
hydrophobic amino acid group with the proviso that at least one of
Y.sub.1 and Y.sub.2 is leucine or isoleucine,
[0025] AA.sub.1, AA.sub.2 and AA.sub.3 independently designate
amino acid groups, wherein the AA.sub.1 and AA.sub.3 amino acid
groups, respectively, may be the same or different when m and n are
more than 1,
[0026] m and n independently represent an integer ranging from 0 to
10,
[0027] a, b and c are independently 0 or 1,
[0028] wherein all the amino acid groups may independently be in a
L, D or DL configuration,
[0029] with the proviso that when X.sub.1 is arginine (R) or lysine
(K), then Y.sub.2 is different from methionine (M).
[0030] By "protecting or blocking group", is in intended to mean
any chemical moiety capable of preventing reactivity of the amino
groups in the peptide, for instance when undergoing a reaction at a
carboxyl group of the molecule. Exemplary protective groups
include, but are not limited to, tBoc (tert-butyloxycarbonyl), Z
(benzoylcarbonyl), Fmoc (fluorenylmethyloxycarbonyl) and Alloc
(allyloxycarbonyl) groups. These and additional protecting and
blocking groups useful in peptide synthesis are well known in the
art.
[0031] Moreover, the term "hydrocarbon group" is intended to mean
any group having carbon and hydrogen atoms in its structure, for
instance in its main chain. This hydrocarbon group may also include
other atoms, such as oxygen and/or nitrogen atoms. Non limiting
examples of R.sub.1 and/or R.sub.2 hydrocarbon groups are alkyl
carbonyl and alkenyl carbonyl groups. Among them, alkyl carbonyl
groups having from 6 to 20 carbon atoms, such as a palmitoyl group,
are preferred. Non limiting examples of R.sub.3 hydrocarbon groups
are alkyl and alkenyl groups comprising from 1 to 20 carbon
atoms.
[0032] Among polar amino acid groups, mention can be made of those
selected from the group consisting of: asparagine (N), serine (S),
cysteine (C), glutamine (Q), aspartic acid (D), glutamic acid (E),
threonine (T), lysine (K), arginine (R), histidine (H), tyrosine
(Y) and tryptophan (W).
[0033] Among small amino acid groups, mention can be made of those
selected from the group consisting of: proline (P), asparagine (N),
aspartic acid (D), threonine (T), valine (V), cysteine (C), alanine
(A), serine (S) and glycine (G).
[0034] Among hydrophobic amino acid groups, mention can be made of
those selected from the group consisting of: alanine (A), glycine
(G), cysteine (C), threonine (T), valine (V), isoleucine (I),
leucine (L), lysine (K), methionine (M), histidine (H), tyrosine
(Y), tryptophan (W) and phenylalanine (F).
[0035] It should be noted that the orientation of the amino- and
c-terminus of the peptide is irrelevant to the inhibitory effect of
the above compounds.
[0036] The groups between brackets in Formula (I) may
advantageously be present (a and/or c equal to 1) so as to
facilitate solubility, membrane penetration and/or detection of the
compounds of this invention or simply for protecting the peptide
against degradation. They can also be useful for strengthening the
binding of these peptides to the AP-2 and/or AP-3 adaptor
complexes.
[0037] In the above formula (I), it is preferred that: [0038]
AA.sub.2 and/or X.sub.4 designates a polar amino acid group which
is preferably charged and more preferably negatively charged,
and/or [0039] X.sub.1 is a charged polar amino acid group, and/or
[0040] X.sub.3 is proline (P), and/or [0041] Y.sub.1 and/or Y.sub.2
and/or Y.sub.3 is an aliphatic amino acid group, such as valine
(V), isoleucine (I) or leucine (L),
[0042] with the proviso that when X.sub.1 is arginine (R) or lysine
(K), then Y.sub.3 is different from methionine (M).
[0043] According to a preferred embodiment of this invention:
[0044] b=1 and AA.sub.2, is glutamic acid (E), [0045] X.sub.1 is
arginine (R), aspartic acid (D) or glutamic acid (E), [0046]
X.sub.2 is glutamine (Q), glutamic acid (E) or asparagine (N),
[0047] X.sub.3 is proline (P), [0048] X.sub.4 is aspartic acid (D)
or glutamic acid (E), [0049] Y.sub.1 is leucine (L), methionine (M)
or isoleucine (I), and preferably leucine (L), [0050] Y.sub.2 is
leucine (L), [0051] Y.sub.3 is valine (V), leucine (L) or
methionine (M), and/or [0052] either n=m=a=c=0, or n=m=c=0 and
a=1,
[0053] with the proviso that when X.sub.1 is arginine (R) or lysine
(K), then Y.sub.3 is different from methionine (M).
[0054] More preferably, all the above conditions are met.
[0055] In a most preferred embodiment of this invention: [0056] b=1
and AA.sub.2 is glutamic acid (E), [0057] X.sub.1 is aspartic acid
(D), [0058] X.sub.2 is glutamic acid (, [0059] X.sub.3; is proline
(P), [0060] X.sub.4 is glutamic acid (E), [0061] Y.sub.1 is leucine
(L), [0062] Y.sub.2 is leucine (L), [0063] Y.sub.3 is methionine
(M), and [0064] either n=m=a=c=0, or n=m=c=0 and a=
[0065] More preferably, in the latter case, Ra.dbd.CH.sub.3,
R.sub.1.dbd.H and R.sub.2.dbd.CO--(CH.sub.2).sub.14--CH.sub.3.
[0066] In another most preferred embodiment: [0067] b=2, [0068]
each of the AA2 groups is glutamic acid (E), and [0069]
Y1=Y2=leucine (L).
[0070] The peptides may be generated by conventional chemical
synthesis or as fusion proteins in protein expression system of any
species or by enzymatic synthesis (Kuliman et al., J. Biol. Chem.,
1980) from constitutive amino acids or theirs derivatives. Peptides
of this invention can still be obtained by biotechnology (use of a
micro-organism, modified or not by genetic engineering); i.e.,
peptides according to this invention can also be obtained by
fermentation of a strain of bacteria, modified or not, by genetic
engineering to produce peptides of sequence previously mentioned
and their fragments. Peptides of this invention can also be
obtained from natural proteins; i.e. by protein extraction of
animal or vegetable origin, followed by controlled hydrolysis which
releases the peptide fragments of average size and of small size,
provided that the released elements contain at least the sequence
included in Formula (I). It is possible, but not necessary, to
extract either the relevant proteins and then to hydrolyze them, or
to initially carry out the hydrolysis on raw extract and then to
purify the peptide fragments. Other more simple or more complex
processes can be considered by the skilled in the art of synthesis,
extraction and purification of proteins and peptides. Thus, the
peptide compounds of this invention may be of natural or synthetic
origin. Preferably, the peptide compounds of this invention are
obtained by chemical synthesis.
[0071] The present invention also pertains to a cosmetic or
dermatological composition comprising at least one peptide compound
as described above, or a mixture thereof, in a physiologically
acceptable medium.
[0072] By "physiologically acceptable medium", it is intended to
designate a carrier adapted for topical application onto skin. This
medium is preferably cosmetically acceptable, i.e. it does not
generate any substantial irritation, redness or heating when
applied onto human skin.
[0073] The amount of compounds of Formula (I) included in this
composition may be any sufficient amount to provide the required
whitening effect. For instance, these compounds may represent from
0.001 to 20% by weight, and more preferably from 0.01 to 10% by
weight, and still more preferably from 0.1 to 5% by weight,
relative to the total weight of the composition.
[0074] This composition can be solid, semi-solid or liquid. It may
be for instance in the form of a powder, ointment, paste, cream,
fluid, milky lotion, cosmetic water, lotion, serum, gel, foam,
facial mask such as a sheet mask, watery or anhydrous stick, and
the like. Preferably, this composition includes water. More
preferably, it is in the form of a gel or of an oil-in-water or
water-in-oil, for instance water-in-silicon, emulsion.
Alternatively, it may be in the form of a multiple emulsion, a
micro-emulsion, nano-emulsion or a dispersion.
[0075] The composition according to this invention may contain
various additives, such as at least one compound chosen from:
[0076] oils, which can be chosen, in particular, from: volatile or
non-volatile, linear or cyclic silicone oils, such as
dimethylpolysiloxanes (dimethicones), polyalkylcyclosiloxanes
(cyclomethicones) and polyalklyphenylsiloxanes
(phenyldimethicones); synthetic oils such as fluorinated oils,
alkyl benzoates and branched hydrocarbons such as polybutene;
vegetable oils and, in particular, soybean or jojoba oil; and
mineral oils such as paraffin oil; [0077] waxes, such as ozokerite,
polyethylene wax, beeswax or carnauba wax; [0078] silicone
elastomers obtained, in particular, by reacting, in the presence of
a catalyst, a polysiloxane having at least one reactive group
(hydrogen or vinyl, in particular) and carrying at least one end
and/or side alkyl (in particular methyl) or phenyl group, with an
organosilicon such as an organohydrogenpolysiloxane; [0079]
surfactants, preferably emulsifiers, whether non-ionic, anionic,
cationic or amphoteric, and, in particular, esters of fatty acids
and polyols, such as esters of fatty acids and glycerol, esters of
fatty acids and sorbitan, esters of fatty acids and polyethylene
glycol; esters of fatty acids and sucrose; esters of fatty alcohols
and polyethylene glycol; alkylpoyglucosides; modified polysiloxanes
polyethers; betaine and its derivatives; polyquaterniums; sulphate
salts of ethoxylated fatty alcohols; sulfosuccinates; sarcosinates;
alkyl- and dialkylphosphates and their salts; and soaps of fatty
acids; [0080] cosurfactants such as linear fatty alcohols and, in
particular, hexadecyl and stearyl alcohols; [0081] thickeners
and/or gelling agents, and, in particular, hydrophilic or
amphiphilic, crosslinked or non-crosslinked homo- and copolymers of
acrylamidoethylpropane sulfonic acid (AMPS) and/or of acrylamide
and/or of acrylic acid and/or of salts or esters of acrylic acid;
xanthan or guar gum; cellulose derivatives; and silicone gums
(dimethiconol); [0082] humectants, such as polyols, including
gylcerin, propylene glycol and sugars, and mucopolysaccharides such
as hyaluronic acid and its salts and esters; [0083] agents for
facilitating percutaneous absorption, such as alcohols, fatty
alcohols and fatty acids and their ester or ether derivatives,
pyrrolidones, terpenes, essential oils and .alpha.-hydroxy acids;
[0084] colorants; [0085] preservatives; [0086] optical modifiers or
soft focus agents such as non-colored and colored, organic and
inorganic materials. Among the materials which may be used are
included: organic pigments, inorganic pigments, polymers and
fillers. The particles that can be present in the present invention
can be natural, synthetic, or semi-synthetic. These optical
modifiers can be platelet-shaped, spherical, elongated or
needle-shaped, or irregularly shaped, surface coated or uncoated,
porous or non-porous, charged or uncharged. Such particles useful
herein include but are not limited to mica, zeolite, kaolin,
silica, boron nitride, lauroyl lysine, nylon, polyethylene, talc,
styrene, polypropylene, polystyrene, ethylene/acrylic acid
copolymer, aluminum oxide, silicone resin, calcium carbonate,
cellulose acetate, PTFE, polymethyl methacrylate, starch. The
particles can be interference pigments with pearl gloss such as
those supplied by EMD Chemicals, Inc. under the trade name TIMIRON
(Registered trademark), COLORONA (Registered trademark) and
supplied by Engelhard Co. under the trade name FLAMENCO (Registered
trademark), TIMICA (Registered trademark). The particles can also
be a composite powder such as a talc/titanium
dioxide/alumina/silica composite powder, for example those sold
under the name Coverleaf AR-(Registered trademark) by the company
Catalyst & Chemicals. Of course, the formulation can contain a
mixture of optical modifiers, each containing characteristics of a
specific visual benefit, to create a combination of visual effects;
[0087] sequestering agents such as the salts of EDTA; [0088]
fragrances; [0089] and their mixtures, without this list being
limiting.
[0090] Examples of such additives and others are cited in
particular in the CTFA Dictionary (International Cosmetic
Ingredient Dictionary and Handbook published by the Cosmetic,
Toiletry and Fragrance Association, 10.sup.th Edition, 2004).
[0091] Furthermore, the topical composition of the present
invention may suitably contain various active agents which may be
chosen from the group consisting of: [0092] antioxidants, such as
ascorbic acid and its derivatives, including ascorbyl palmitate,
ascorbyl tetraisopalmitate, ascorbyl glucoside, magnesium ascorbyl
phosphate, sodium ascorbyl phosphate and ascorbyl sorbate;
tocopherol and its derivatives, such as tocopheryl acetate,
tocopheryl sorbate and others esters of tocopherol; BHT and BHA;
and plant extracts, for instance from Chondrus cripsus, Rhodiola,
Thermus thermophilus, mate leaf, oak wood, kayu rapet bark, sakura
leaves and ylang ylang leaves; [0093] anti-ageing agents, such as
acyl aminoacids (for instance Maxilip, Matrixyl 3000 or Biopeptide
CL from SEDERMA or Sepilift from SEPPIC), Pisum sativum extracts,
hydrolyzed soy proteins, methylsilanol derivatives such as
methylsilanol mannuronate, hydrolyzed cucurbita pepo seedcake,
Scenedesmus extract; [0094] anti-pollution agents such as Moring a
pterygosperma seed extracts; [0095] keratolytic agents, such as
.alpha.-hydroxyacids (for instance, glycolic, lactic, citric,
malic, mandelic or tartaric acid) and .beta.-hydroxyacids (for
instance, salicylic acid), and their esters, including C.sub.12-13
alkyl lactate, and plant extracts containing these hydroxyacids,
such as Hibiscus sabdriffa extracts; [0096] astringents such as
hamamelis extracts; [0097] moisturizers, including plant extracts
such as Castanea sativa extracts, hydrolyzed hazelnut proteins,
Polyanthes tuberosa polysaccharides, Argania spinosa kernel oil and
an extract of pearl containing conchiolin that is sold especially
by the company Maruzen (Japan) under the trade name Pearl
Extract.RTM.; homo- and copolymers of
2-metharyloyloxyethylphosphorylcholine, such as Lipidure HM and
Lipidure PBM from NOF; saccharides such as glucose, fructose,
mannose or trehalose; glycosaminoglycanes and their derivatives
such as hyaluronic acid, sodium hyaluronate and acetylated
hyaluronic acid; panthenol; allantoin; aloe vera; free amino acids
and their derivatives; glucosamine; citric acid; urea and its
derivatives and ceramides; [0098] emollients such as glyceryl
polymethacrylate; anti-inflammatory agents, such as bisabolol,
allantoin, tranexamic acid, zinc oxide, sulfur oxide and its
derivatives, chondroitin sulfate, glycyrrhizinic acid and its
derivatives such as glycyrrhizinates; [0099] and their
mixtures.
[0100] The topical composition can also include organic and/or
inorganic sunscreens. Among organic sunscreens, mention can be made
of dibenzoylmethane derivatives such as butyl
methoxydibenzoylmethane (Parsol 1789 from HOFFMANN LA ROCHE),
cinnamic acid derivatives such as ethylhexyl methoxycinnamate
(Parsol MCX from HOFFMANN LA ROCHE), salicylates, para-aminobenzoic
acids, .beta.- .beta.'-diphenylacrylate derivatives, benzophenone
derivatives, benzylidenecamphor derivatives such as
terephtalylidene dicamphor sulphonic acid, phenylbenzimidazole
derivatives, triazine derivatives, phenylbenzotriazole derivatives,
anthranilic derivatives, all of which may be coated or
encapsulated. Among inorganic photoprotective agents, mention can
be made of pigments or alternatively nanopigments formed from
coated or uncoated metal oxides, such as, for example, titanium
oxide, iron oxide, zinc oxide, zirconium oxide or cerium oxide
nanopigments; which are all CV photoprotective agents well known
per se.
[0101] In addition, the pH of the topical composition of the
present invention is preferably in the range from 4 to 8, and
preferably from 4.5 to 7.
[0102] The cosmetic compositions according to the present invention
may composition according to the invention contains at least one
whitening agent able to block the synthesis of structural proteins
involved in the mechanism of melanogenesis (stage I) such as the
melanocyte-specific glycoprotein Pmel17. Such an active agent may
be the tranexamic cetyl ester
(trans-4(aminomethyl)cyclohexanecarboxylic acid hexadecyl ester
hydrochloride) sold by Nikko Chemicals (Japan) or the ferulic acid
Cytovector (water, glycol, lecithin, ferulic acid,
hydroxyethylcellulose) sold by BASF under the trade name
Cytovector.RTM..
[0103] As a variant or in addition, the composition according to
the invention may comprise a whitening agent having an inhibition
effect on melanin synthesis and/or an inhibition effect on MITF
expression and/or an anti-tyrosinase activity and/or an inhibition
effect on endothelin-1 synthesis, such as a licorice extract
(glycyrrhiza glabra extract), which is sold especially by the
company Maruzen under the trade name Licorice Extract.RTM..
[0104] As a variant or in addition, the composition according to
the invention may comprise a whitening agent having an antioxidant
effect as well, such as vitamin C compounds, including ascorbate
salts, ascorbyl esters of fatty acids or of sorbic acid, and other
ascorbic acid derivatives, for example, ascorbyl phosphates, such
as magnesium ascorbyl phosphate and sodium ascorbyl phosphate, or
saccharide esters of ascorbic acid, which include for instance
ascorbyl-2-glucoside, 2-O-- alpha-D-glucopyranosyl L-ascorbate, or
6-O-- beta-D-galactopyranosyl L-ascorbate. An active agent of this
type is sold especially by the company DKSH under the trade name
Ascorbyl Glucoside.RTM..
[0105] Other whitening agents may be included in the compositions
according to this invention. Mention can be made of depigmenting
agents such as plant extracts including Narcissus tazetta extracts;
arbutin; kojic acid; ellagic acid; cysteine; 4-thioresorcin;
resorcinol or rucinol or their derivatives; glycyrrhizinic acid and
hydroquinone-beta-glucoside.
[0106] These compositions according to this invention may be used
for depigmenting human skin in case of irregular pigmentation
pattern due to acquired hyperpigmentation such as melasma
(chloasma); postinflammatory melanoderma; solar lentigo; age spots
(lentigo senile); pigmentation spots that appear on the skin upon
sun exposure often in conjunction with drugs such as birth control
pill or other hormonal medication, or following the application of
a perfume, or during pregnancy; discoloration due to chemical peels
and dermabrasion, pre- and post-laser resurfacing, or pre- and
post-laser hair removal; pigmented keratosis or hypopigmentation
after traumas (scars). Moreover they may also be used for
lightening/brightening the complexion or forms of hyperpigmentation
and hypopigmentation mentioned above.
[0107] The present invention thus also pertains to a cosmetic
method for whitening, bleaching or lightening human skin,
comprising topically applying onto human skin a cosmetic
composition as described above.
[0108] It also relates to the use of the above composition for
manufacturing a dermatological preparation intended to depigment
human skin.
[0109] The skin areas on which the above process and use may be
carried out may be any region of human skin preferably excluding
the scalp, such as facial skin, breast skin, hand and arm skin or
leg skin.
[0110] This invention will be better understood by reference to the
following non-limiting Examples, taken in combination with the
attached drawings in which:
[0111] FIG. 1 is a histogram showing the inhibiting effect of
various compounds of this invention on the binding of AP-2 to the
tyrosine-based signal containing TGN38 peptide;
[0112] FIG. 2 is a histogram showing the inhibiting effect of
various compounds of this invention on the binding of AP-3 to the
tyrosine-based signal containing Lamp-1 peptide;
[0113] FIG. 3 is a histogram showing the inhibiting effect of
various compounds of this invention on the binding of AP-2 to the
dileucine-based sorting signal containing tyrosinase; and
[0114] FIG. 4 is a histogram showing the inhibiting effect of
various compounds of this invention on the binding of AP-3 to the
dileucine-based sorting signal containing tyrosinase.
EXAMPLES
[0115] The following examples further illustrate embodiments within
the scope of the present invention. The examples are given solely
for the purpose of illustration and are not to be construed as
limitations of the present invention, as many variations thereof
are possible without departing from the scope of the invention.
Example 1
Binding Test--Study of the Interactions of the Compounds According
to this Invention with AP-2 and AP-3 Adaptor Complexes
[0116] Synthesis:
[0117] Five peptide compounds in accordance with this invention
were synthesized using amino acids protected with Fmoc
[(N-(9-fluorenyl)methoxycarbonyl) and activated with
benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluoro-phosphate
and a peptide synthesizer. After cleavage from the resin and the
protecting groups, peptides were purified by reverse phase HPLC
using Delta Pac C-18 columns (Millipore) and an elution from 0-50%
acetonitrile in 0.1% trifluoroacetic, water for 50 min. Purity of
all peptides were 90% or better and was confirmed by HPLC, UV
spectrometry, and mass spectrometry. All peptides were freeze dried
and stored at -20.degree. C. until use. Prior to biosensor
experiments, all peptides were dissolved in HPLC-pure H.sub.2O to a
stock concentration of 5 mM. Aliquots of this stock were frozen to
avoid repeated freeze/thawing.
[0118] The compounds thus made consisted in the following peptide
sequences:
TABLE-US-00001 Compound 1: DERQPLLVE Compound 2: EERQPMLLD Compound
3: AEDEPLLME Compound 4: AEDEPLLVD Compound 5: MVEENPILME
[0119] None of these peptides is found in any known human protein.
These compounds were tested for their ability to block the binding
of adaptor complexes such as AP-2 and AP-3, which are key
components in the melanogenesis process, to the human tyrosinase
tail.
[0120] Test Protocol:
[0121] It is well established that adaptors such as AP-2 and AP-3
bind sorting signal containing peptides. Among them, the
tyrosine-based signal of TGN38 is the most efficient AP-2 binding
sequence known so far, while the same type of signal from Lamp-1
binds AP-1 and AP-3 and thus serves as a control for AP-3 binding
in our experiments.
[0122] Human tyrosinase also contains tyrosine residues in its
cytoplasmic tail sequence, however they are not relevant with
respect to adaptor binding. Instead, the tail contains a
dileucine-based sorting signal that is bound by adaptors such as
AP-2 and AP-3.
[0123] Binding between adaptors and the tyrosinase tail peptide was
recorded in real-time using a BIAcore 3000 biosensor (GEHealthcare)
based on surface plasmon resonance. The tyrosinase tail peptide
(sequence CRHKRKQLPEEKULLMEKEDYHSLYSHL) was immobilized on the
surface of a CM5 sensor surface using thiol coupling and resulting
in the immobilization of .ltoreq.300 RU of peptide. In brief, the
CM5 surface was activated with EDC/NHS for 2 min at a flow-rate of
10 .mu.l/min, followed by modification with PDEA for another 2 min.
Subsequently, peptide was injected at a flow-rate of 5 .mu.l/min
for 1 min at a concentration of 5 .mu.g/ml in 10 mM sodium acetate
pH 4.5. The flow-rate was then adjusted to 20 .mu.l/min and
remaining active groups were blocked by injection of 50 mM
cysteine, 1M NaCl. Following peptide immobilization, the surface
was washed with buffer A (50 mM Tris pH 8.0, 250 mM NaCl, 0.005%
Tween-20) at a flow rate of 30 .mu.l/min. Purified adaptors were
then injected at concentrations ranging from 100-1000 nM in buffer
A for 2 min followed by dissociation for 2 min. The surface was
regenerated by 2 pulse injection of 20 sec with 50 mM NaOH and 10
mM NaOH, 0.5% SDS. As controls for the sequence specific binding of
the AP-2 and AP-3 adaptors, the tyrosine-based sorting signal
peptides of TGN 38 (sequence--CRPKASDYQRL) and Lamp-1
(sequence--CRKRSHAGYQTI) were immobilized to the sensor surface
exactly as described for the tyrosinase peptide.
[0124] For competition experiments, 200 nM adaptors were incubated
with a 10-1000 fold molar excess of one of the 5 compound peptides
for 10 min prior to injection. For comparison of adaptor binding to
the immobilized tyrosinase tail, the RU values at the end of the
injection (association period) and after the dissociation period
were evaluated.
[0125] Results:
[0126] The results of these experiments are shown on the attached
drawings.
[0127] As illustrated in FIG. 1, the five compounds of this
invention did not significantly inhibit the binding of AP-2 to
TGN38, compared to RPKASDYQRL which was used as a positive control
since this peptide is known to bind to the .mu.2 chain of the AP-2
heterotetramer. It can thus be derived that the test compounds do
not bind to the .mu.2 chain of AP-2.
[0128] Moreover, as illustrated in FIG. 2, the five compounds of
this invention did not significantly inhibit the binding of AP-3 to
Lamp-1, compared to RKRSHAGYQTI which was used as a positive
control since this peptide is known to bind to the .mu.3 chain of
the AP-3 heterotetramer. It can thus be derived that the test
compounds do not bind to the .mu.3 chain of AP-3.
Example 2
Binding Test--Study of the Inhibiting Effect of the Compounds
According to this Invention on Tyrosinase Binding to AP-2 and
AP-3
[0129] A test similar to that described in Example 1 was conducted
to determine the effect of the five compounds synthesized in
Example 1 on the binding of tyrosinase to AP-2 and AP-3. The molar
amount of adaptor and soluble peptides used for inhibition were
exactly the same as outlined in Example 1.
[0130] The same control sequences were used as those used in
Example 1 as negative controls since it is known that tyrosinase
does not bind to the .mu.2 and .mu.3 chains of AP-2 and AP-3,
respectively. Moreover, EEKQPLLME was used as a positive control,
since this sequence harbors the part of human tyrosinase that is
known to bind to AP-2 and AP-3.
[0131] As can be seen in FIG. 3, the five compounds of this
invention drastically inhibited the binding of AP-2 to tyrosinase
with compound 4 being the most active.
[0132] Moreover, as shown in FIG. 4, these compounds also very
efficiently inhibited the binding of AP-3 to tyrosinase, with
compound 4 again being the most active.
[0133] These examples thus demonstrate that the compounds according
to this invention effectively block the binding of AP-2 and AP-3 to
tyrosinase. It is thus believed that they will block the correct
intracellular sorting of tyrosinase to melanosomes and thus melanin
synthesis.
Example 3
Study of the Melanocyte Intracellular Penetration of the Peptide
According to this Invention
[0134] Test Protocol:
[0135] B16 melanoma cells were cultured at 37.degree. C. and 5% of
CO.sub.2 in microplates with 96 wells in the presence of DMEM
(Invitrogen, reference 11880028) containing 1 g/l of glucose
without phenol red supplemented by 3 g/l of glucose (Sigma,
reference G7021), 2 mM of L-glutamine, 50 UI/ml of penicillin, 50
.mu.g/ml of streptavidin (Invitrogen, reference 15070063) and 10%
of fetal veal serum (Invitrogen, 10270098). After 24 h, the culture
medium is replaced by a DMEM medium containing or not (control) a
stable derivative of .alpha.-MSH, i.e. NDP-MSH (NLE-4-D-PHE-7-A
Melanocytes Stimulating Hormone, Sigma, reference M-8764) and
containing or not the peptide (compound 3 of example 1 or its
N-palmitoyl derivative) according to the invention. Each peptide
was tested with and without lipofectamine, (Invitrogen, reference
1538-100), an agent used to facilitate the peptide penetration. The
melanocytes were then incubated for 72 hours at 37.degree. C. and
5% of CO.sub.2.
[0136] At the end of the incubation period, the medium was
eliminated and cells were rinsed with a solution of PBS (phosphate
buffer saline). The penetration of each peptide was analyzed by
microscopy (In Cell Analyser.RTM. 100, GE Healthcare) with the
objective .times.20 or by Flux Cytometry after unsticking of cells
by enzymatic treatment with trypsine (Flux FACSarray, Becton
Dickinson).
[0137] Results:
[0138] The following table 1 illustrates the measurement of the
penetration of the peptide+FITC (fluoresceine isothiocyanate) in
B16 melanocytes at different peptide concentrations (4, 20, 100 and
500 .mu.g/ml).
TABLE-US-00002 TABLE 1 Control without Compound 3 according to
Example 1 lipofectamine without lipofectamine (% control) (%
control) 4 .mu.g/ml 20 .mu.g/ml 100 .mu.g/ml 500 .mu.g/ml 100% ns *
263 533 1640 Compound 3 according to Example 1 Control with with
lipofectamine (% control) lipofectamine 4 .mu.g/ml 20 .mu.g/ml 100
.mu.g/ml 500 .mu.g/ml 109 166 338 1097 4265 N-palmitoyl derivatives
of compound Control without 3 according to Example 1 lipofectamine
without lipofectamine (% control) (% control) 4 .mu.g/ml 20
.mu.g/ml 100 .mu.g/ml 500 .mu.g/ml 100 276 364 1486 982 N-palmitoyl
derivatives of compound Control with 3 according to Example 1
lipofectamine with lipofectamine (% control) (% control) 4 .mu.g/ml
20 .mu.g/ml 100 .mu.g/ml 500 .mu.g/ml 120 765 1495 1331 7726 * ns:
p > 0.05, not significant
[0139] The observation in fluorescence microscopy clearly
demonstrates that peptides penetrate into cells and that this
penetration increases with the addition of lipofectamine. These
results were confirmed by flux cytometry, which showed that the
intensity of fluorescence depends directly on the peptide
concentration.
Example 4
Effect of the N-Palmitoyl Derivative of Compound 3 of Example 1 on
Melanogenesis
[0140] Test Protocol:
[0141] The protocol used is the same as in example 3. The
melanocytes were incubated for 96 hours at 37.degree. C. and 5% of
CO.sub.2. Kojic acid was used as a positive control.
[0142] At the end of the incubation period, the quantity of
melanine was evaluated by measuring the absorption at 405 nm for
each sample.
[0143] Results:
[0144] The following table 2 illustrates the effect on the
synthesis of melanine of the N-palmitoyl derivative of compound 3
of example 1 at 0.5; 1; 2.5; 5 and 10 .mu.g/ml.
TABLE-US-00003 TABLE 2 N-palmitoyl derivative of compound 3 Control
without of example 1 with out lipofectamine, lipofectamine, with
NDP-MSH (% of inhibition) with NDP-MSH 0.5 1 2.5 5 10 (% of
inhibition) .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml .mu.g/ml 0 ns * 9
10 14 20 Kojic acid with NDP-MSH Control (% of inhibition) with
NDP-MSH 6.9 15.6 36 80 (% of inhibition) .mu.g/ml .mu.g/ml .mu.g/ml
.mu.g/ml 0 -13 -7 33 75 * ns: p > 0.05, not significant
[0145] The presence of NDP-MSH sharply inferred the melanogenesis.
This induction of the synthesis of melanin was sharply inhibited by
kojic acid tested between 36 and 80 .mu.g/ml. These results
validate the test.
[0146] The presence of lipofectamine did not modify the synthesis
of melanin by B16 melanocytes incubated with NDP-MSH.
[0147] The treatment with the N-palmitoyl derivative of compound 3
of example 1, tested between 1 and 10 .mu.g/ml, in the absence of
lipofectamine, allowed to inhibit sharply the melanogenesis boosted
by the NDP-MSH (from 9 to 20% of inhibition). This inhibition
increased with the presence of lipofectamine (from 5 to 29% of
inhibition between 1 to 10 .mu.g/ml).
[0148] Thus, this study confirms that peptides according to the
invention, such as the N-palmitoyl derivative of compound 3 of
example 1, inhibit the synthesis of melanine by the
melanocytes.
Example 5
Cosmetic Compositions
[0149] The compositions given hereafter may be prepared by
conventional methods from the following components. The contents
are expressed by weight relative to the total weight of the
composition.
TABLE-US-00004 Formula 5.1: Water in oil formula INCI NAME QUANTITY
Ethylhexyl palmitate 14.00 Propylparaben 0.30 PEG-30
dipolyhydroxystearate 3.00 Silica dimethyl silylate 2.00
Hydrogenated polyisobutene 31.49 Ethylene/propylene/styrene
copolymer 2.62 Butylene/ethylene/styrene copolymer 0.88 BHT 0.01
Water 42.10 Sodium chloride 0.80 Tetrasodium EDTA 0.05 Glycerin
2.00 Methylparaben 0.30 Xanthan gum 0.35 Peptide according to the
invention - Compound 3 0.10
TABLE-US-00005 Formula 5.2: Serum INCI NAME QUANTITY Water 85.19
Tetrasodium EDTA 0.05 Polyethyleneglycol 5.00 Acrylates/C.sub.10-30
alkyl acrylate crosspolymer 0.35 Ammonium
acryloyldimethyltaurate/VP copolymer 0.30 Glycerin 4.43 PEG-8 1.00
Sodium polyacrylate 0.04 Caprylyl glycol 0.15 PEG-11 methyl ether
dimethicone 3.00 Propylene glycol 0.01 Propylparaben 0.01
Methylparaben 0.31 Sodium hydroxide 0.06 Peptide according to the
invention - N-palmitoyl 0.10 derivative of Compound 3
TABLE-US-00006 Formula 5.3: Gel-cream formula INCI NAME QUANTITY
Water 75.36 Tetrasodium EDTA 0.05 Glycerin 7.26 Ammonium
acryloyldimethyltaurate/VP copolymer 0.80 Acrylates/C.sub.10-30
alkyl acrylate crosspolymer 0.15 Methylparaben 0.30 Cyclomethicone
8.05 Polysorbate-20 0.05 Isononyl isonanoate 3.00 Dimethicone 2.00
Phenyl trimethicone 2.00 Tocopheryl acetate 0.50 Polyacryl amide
0.12 Hydrogenated polyisobutene 0.05 Laureth-7 0.02 Hydroxyethyl
acrylate/sodium acryloyldimethyl 0.09 taurate copolymer Squalane
0.01 Polysorbate 60 0.01 Sorbitan isostearate 0.02 Sodium hydroxide
0.02 Peptide according to the invention - Compound 5 0.10
Sequence CWU 1
1
1119PRTArtificialWHITENING AGENT 1Asp Glu Arg Gln Pro Leu Leu Val
Glu1 529PRTArtificialWHITENING AGENT 2Glu Glu Arg Gln Pro Met Leu
Leu Asp1 539PRTArtificialWHITENING AGENT 3Ala Glu Asp Glu Pro Leu
Leu Met Glu1 549PRTArtificialWHITENING AGENT 4Ala Glu Asp Glu Pro
Leu Leu Val Asp1 5510PRTArtificialWHITENING AGENT 5Met Val Glu Glu
Asn Pro Ile Leu Met Glu1 5 10630PRTArtificialTyrosinase tail
peptide 6Cys Arg His Lys Arg Lys Gln Leu Pro Glu Glu Lys Gln Pro
Leu Leu1 5 10 15Met Glu Lys Glu Asp Tyr His Ser Leu Tyr Gln Ser His
Leu 20 25 30711PRTArtificialTGN 38 fragment 7Cys Arg Pro Lys Ala
Ser Asp Tyr Gln Arg Leu1 5 10812PRTArtificiallamp-1 fragment 8Cys
Arg Lys Arg Ser His Ala Gly Tyr Gln Thr Ile1 5
10910PRTArtificialcontrol 9Arg Pro Lys Ala Ser Asp Tyr Gln Arg Leu1
5 101011PRTArtificialcontrol 10Arg Lys Arg Ser His Ala Gly Tyr Gln
Thr Ile1 5 10119PRTArtificialcontrol 11Glu Glu Lys Gln Pro Leu Leu
Met Glu1 5
* * * * *